1,4-dioxane-2,5-dione

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1999-11-11 to 1999-12-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

2000

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

1984

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Glycollic acid

EC Number:

201-180-5

EC Name:

Glycollic acid

Cas Number:

79-14-1

Molecular formula:

C2H4O3

IUPAC Name:

glycol acid

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution of 800 mg/L was prepared by weighing approximately 1.6 g of H-24277, bringing this up to 2000 mL in well water and stirring for 30 minutes. Nominal concentrations of test substance solutions and stock solutions were not adjusted for purity. Test solutions of 25, 50, 100, 200, 400, and 800 mg/L were prepared by adding the appropriate volume of stock solution for a total test solution volume of 1-liter.
- Controls: dilution water control
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain/clone: Daphnia magna
- Age of parental stock (mean and range, SD): 28 days
- Feeding during test: no


ACCLIMATION
- Type and amount of food: During acclimatiion Daphnids were fed on a daily basis with a yeast, cereal leaves and trout chow (YCT) mixture (standardized to 1800 mg/L total solids) and the green alga, Selenastrum capricornutum at a rate of 62,500 cells/mL (the combination of YCT and alga is equivalent to 0.1-0.2 mg total organic carbon per daphnid).
- Feeding frequency: daily
- Health during acclimation (any mortality observed): Sickness, injury, and abnormalities were not seen and ephippia were not being produced by the parent daphnids. No adult immobility was seen in the cultures used for testing during the 48-hour pretest period.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

120 to 123 mg/L CaCO 3

Test temperature:

20.4 to 20.7 °C

pH:

At high concentrations: 200, 400 and 800 mg/L the pH ranged between 3.0 and 3.8, at lower concentrations: 25, 50 and 100 mg/L the pH ranged from 6.9 to 7.7

Dissolved oxygen:

8.9 mg/L

Salinity:

n.a.

Conductivity:

290 to 490 µmhos/cm

Nominal and measured concentrations:

nominal concentrations: 0, 25, 50, 100, 200, 400 and 800 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Pyrex ® beakers
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Pyrex®, 250 mL fill volume, 200 mL solution volume
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Dilution water originated from the Haskell Laboratory well which is 378-feet deep and is cased and sealed into bedrock to prevent contamination. The hardness of the well water is adjusted to approximately 110-140 mg/L as CaCO 3 by the flow-proportioned addition of CaCl 2. The well water is then aerated, passed through a green sand filter to remove iron, and filtered through 50-, 10-, and 5-µm filters to remove particulates. The water is heated or chilled as appropriate and distributed through aged polyvinyl chloride piping.
- Total organic carbon: 0.30
- Metals: ND
- Pesticides: ND
- Chlorine: 0.04
- Alkalinity: 54 mg/L CaCO 3 at test start
- Ca/mg ratio: 14.1
- Conductivity: 490 µmhos/cm at 800 mg/L
- Culture medium different from test medium: no
- Intervals of water quality measurement: every two years

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light (approximately 510 - 521 Lux) and 8 hours darkness was employed which included 30 minutes of transitional light (7 - 8 Lux)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Test concentrations: An initial study was conducted at nominal test concentrations of 6.3, 12.5, 25, 50, and 100 mg/L (5 daphnids per replicate, 4 replicates per concentration). A dilution water control was used in the study.
- Results used to determine the conditions for the definitive study: The only immobility recorded at 48 h was 5% at the 12.5 mg/L nominal test concentration, therefore, the definitive test concentrations were 25, 50, 100, 200, 400 and 800 mg/L.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

141 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Reported statistics and error estimates:

The 48-hour EC 50 and 95% confidence intervals were calculated by the binomial method based on nominal concentrations. The highest nominal concentration causing no immobility at test end and the lowest nominal concentration causing 100% immobility at test end were assessed by visual observation.

Any other information on results incl. tables

Table 1: Effect of glycolic acid on Immobilisation of Daphnia magna

Treatment (mg a.i./L) [nominal conc. used]		Number of immobile Daphnids/Number of Daphnids at Test Start
		[h] 24		[h] 48
		Cumul. no. affected	% affected	Cumul. no. affected	% affected
Control (dilution water only)	1	0/5	0	0/5	0
	2	0/5	0	0/5	0
	3	0/5	0	0/5	0
	4	0/5	0	0/5	0
25	1	0/5	0	0/5	0
	2	0/5^2d	40	0/5	0
	3	0/5^3d	60	0/5	0
	4	0/5^1d	20	0/5	0
50	1	0/5^1d	20	0/5	0
	2	0/5^3d	60	0/5	0
	3	0/5	0	0/5	0
	4	0/5	0	0/5	0
100	1	0/5^1d	20	0/5	0
	2	0/5^1d	20	0/5	0
	3	0/5	0	0/5	0
	4	0/5	0	0/5	0
200	1	5/5	100	5/5	100
	2	5/5	100	5/5	100
	3	5/5	100	5/5	100
	4	5/5	100	5/5	100
400	1	5/5	100	5/5	100
	2	5/5	100	5/5	100
	3	5/5	100	5/5	100
	4	5/5	100	5/5	100
800	1	5/5	100	5/5	100
	2	5/5	100	5/5	100
	3	5/5	100	5/5	100
	4	5/5	100	5/5	100

 OBSERVATION KEY

a Daphnid lethargic;b Daphnid visibly small in size;c Daphnid pale in color;d Daphnid floating at surface;e Daphnid accidentally crushed by pipette during transfer

Applicant's summary and conclusion

Validity criteria fulfilled:

not specified

Conclusions:

The present study was performed according to OECD Test Guideline 202 in order to evaluate the 48–hr-acute toxicity of glycolic acid to Daphnia magna under static conditions. Daphnids were exposed to control and test chemical at nominal concentration of 25, 50, 100, 200, 400 and 800 mg a.i./L for 48 hr. The 48– hour LC50/EC50 was 141 mg a.i./L. The described effects are considered to be based on a decrease of pH in the incubation medium caused by glycolic acid, the measured initial pH of the incubation media were: 7.2 in the solution with 25 mg/L, 7.1 at 50 mg/L, 6.9 at 100 mg/L, 3.8 at 200 mg/L, 3.3 at 400 mg/L and 3.1 at 800 mg/L. After 48h the pH in the 25, 50 and 100 mg/L group returned to physiological values. Thus, the observed effects are supposed to be related to acidity.